A green and easy-to-assemble electrochemical biosensor based on thylakoid membranes for photosynthetic herbicides detection

In this study, we report on an easy-to-assemble amperometric electrochemical biosensor incorporating thylakoid membranes for the detection of photosynthetic herbicides. These molecules interfere with the light-induced photosynthetic electron transport occurring at the level of the photosystems within the thylakoid membranes, thus reducing the current of the associated bioelectrode. Thylakoid membranes isolated from pea plants were adsorbed directly on a bare carbon paper working electrode and placed in the measurement cell in the absence of any electrochemical mediator, obtaining a fully environmental-friendly biodevice capable of photocurrent densities up to 14 μA/cm². Three photosynthetic herbicides inhibiting Photosystem II and belonging to different chemical classes, namely diuron, terbuthylazine and metribuzin, were detected by measuring the electrode photocurrent, which decreased reproducibly in a concentration-dependent manner in a range between 10⁻⁷ – 5 × 10⁻⁵ M of each herbicide. The limit of detection for the three herbicides was between 4–6 × 10⁻⁷ M. Storage stability tests revealed for the biosensor a half-life longer than 15 days at 4 °C and full stability up to 4 months at −80 °C. This study provides a simple, environmental-friendly and cost-effective procedure for the fabrication of a mediatorless carbon paper-based electrochemical biosensor characterized by high photocurrents, long storage stability, reproducible detections and good sensitivity.